Becton Dickinson and Company Limited

James Ghadiali

Jody Martin

Eleen Shum

Christina Chang

Christina Fan

Katherine Lazaruk

Gretchen Yinbon Lam

Nidhanjali Bansal

Robert J Radford

Adam Wright

Brent S Gaylord

Douglas Byron Sherman

Dennis E Prosen

Olaf Zoellner

Jason G Vidal

Eric Jensen

Devon Joseph Jensen

Mirko Corselli

David Rosenfeld

Paul Waterman

Janice H Lai

Jurg Rohrer

Aaron Jacob Tyznik

Devon Jensen

Disclosed herein include methods, compositions, and kits suitable for spectral unmixing and compensation in multi-parameter flow cytometry. Disclosed herein include reagents which comprise two antibodies conjugated to one another to form a bispecific reagent (e.g., a bispecific probe). A first antibody can have affinity for a highly expressed antigen on the surface of a cell of interest and a second antibody can have affinity for each of the antibody-dye conjugates in a multi-parameter panel. There are provided, in some embodiments, methods of using the bispecific reagent to determine spillover, perform compensation, and generate a compensation matrix.

Bi-specific probes to enable the use of single-cell samples as single color compensation control

Disclosed herein include systems, methods, compositions, and kits for sample identification. A sample indexing composition can comprise, for example, a protein binding reagent associated with a sample indexing oligonucleotide. Different sample indexing compositions can include sample indexing oligonucleotides with different sequences. Sample origin of cells can be identified based on the sequences of the sample indexing oligonucleotides. Sample indexing oligonucleotides can be barcoded using barcoded and lengthened using daisy-chaining primers.

Sample indexing for single cells

Disclosed herein include systems, methods, compositions, and kits for determining protein expression and gene expression simultaneously and for sample indexing. In some embodiments, an oligonucleotide associated with a cellular component-binding reagent (e.g., an antibody) comprises one or more of a unique molecular label sequence, a primer adapter, antibody-specific barcode sequence, an alignment sequence, and/or a poly(A) sequence. In some embodiments, the oligonucleotide is associated with the cellular component-binding reagent via a linker (e.g., 5AmMC12).

Oligonucleotides associated with antibodies

Some embodiments disclosed herein provide a plurality of compositions each comprising a protein binding reagent conjugated with an oligonucleotide. The oligonucleotide comprises a unique identifier for the protein binding reagent it is conjugated with, and the protein binding reagent is capable of specifically binding to a protein target. Further disclosed are methods and kits for quantitative analysis of a plurality of protein targets in a sample and for simultaneous quantitative analysis of protein and nucleic acid targets in a sample. Also disclosed herein are systems and methods for preparing a labeled biomolecule reagent, including a labeled biomolecule agent comprising a protein binding reagent conjugated with an oligonucleotide.

Measurement of protein expression using reagents with barcoded oligonucleotide sequences

Aspects of the present disclosure include systems for use in preparing an epitope tagged biomolecule reagent. A reagent preparatory apparatus for preparing the epitope tagged biomolecule reagent from an activated biomolecule and activated epitope tag is also described. Methods for communicating and receiving an epitope tagged biomolecule reagent request and preparing the subject epitope tagged biomolecule reagents are also provided.

Methods and systems for providing epitope tagged biomolecules

Fluorogenic sensors for a target analyte are provided. The fluorogenic sensors can include a water soluble light harvesting multichromophore and a fluorogenic dye covalently linked to the multichromophore and in energy-receiving proximity therewith. Aspects of the fluorogenic sensors include selective recognition of the target analyte by chemical reaction or by specific binding. In some cases, the linked fluorogenic dye includes a lactone or spiro-lactone group that is configured to chemically react with a target analyte via lactone ring-opening to produce a fluorescent dye configured for excitation by the multichromophore. Also provided are labelled specific binding members that include the subject dyes. Methods of evaluating a sample for the presence of a target analyte and methods of labelling a target molecule in which the subject fluorogenic sensors find use are provided. Systems, devices and kits for practicing the subject methods are provided.

Polymeric dye ratiometric sensor for analyte detection and methods of using the same

Photostable polymeric tandem dyes are provided. The photostable polymeric tandem dyes include a water soluble light harvesting multichromophore and a luminescent metal complex covalently linked to the multichromophore and in energy-receiving proximity therewith. Also provided are labelled specific binding members that include the subject dyes. Methods of evaluating a sample for the presence of a target analyte and methods of labelling a target molecule in which the subject polymeric tandem dyes find use are also provided. Systems and kits for practicing the subject methods are also provided.

Photostable fluorescent polymeric tandem dyes including luminescent metal complexes

Methods for producing a high resolution image of a sample are provided. In some embodiments, the method includes: detecting first and second sets of spatially-dependent emission signals from a sample labeled with a fluorescent polymeric dye; and producing a high resolution fluorescence image of the sample from the detected first and second sets of spatially-dependent emission signals. In some embodiments, the sample is a cell. Also provided are systems for imaging a sample that include a high resolution light microscope including a light source configured to irradiate a field of view with an excitation light; a photodetector configured to detect an emission signal: and a polarization modulator disposed in the light pathway between the light source and the photodetector; and a sample labelled with a polymeric dye and disposed in the field of view.

Methods and systems for high resolution fluorescence microscopy of polymeric dye-labeled samples using polarized light

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San Diego, CA, United States of America

James Ghadiali